ABSTRACT
Objective To study the expression and biological functions of RN181 in SMMC7721 cells,the retroviral vector was constructed.Methods Gene cloning techniques were used to construct pRetrox-TRE3G/RN1S1 recombinant vector.The regulating plasmid pRetroX-TRE3G/RN181 or the response plasmid of pRetroX-Tet3G were respectively cotansfected into GP2-293 cells with Envelope Vector plasmid to package retrovirus after routine identification.Both viruses co-infected target cells SMCC7721,and then were selected by G418 to obtain stable cell lines.The stable cell lines were induced by doxycycline (DOX),and then verified by RT-PCR and Western blotting.CCK-8 was used to evaluate the effect of RN181 on growth of SMMC7721 cells.Results We constructed the recombinant plasmid.Stable recombinant plasmid were verified by screening.And there were significant differences of RN181 between the induced and uninduced cell lines through RT-PCR and Western blot.Conclusions We have successfully constructed the inducible stable RN181 expression SMCC7721 cell,which can be used as an effective cell model to study the biological functions of RN181.We found RN181 could suppress the proliferation and invasion in SMMC7721 cells in vitro.
ABSTRACT
Objective To study the clinical results of patients with gallbladder stones and common bile duct stones treated either by laparoscopic cholecystectomy followed by endoscopic sphincterotomy (EST + LC) or laparoscopic cholecystectomy + common bile duct exploration (LC + LCBDE) in the elderly patients.Methods A retrospective study was conducted on 96 patients who had common bile duct and gallbladder stones treated from January 2012 to January 2016.The patients were divided into the control group and the observation group.46 patients were in the observation group who underwent LC three days after EST,while the remaining patients were in the control group who underwent LC and LCBDE.The serum amylase levels before LC,operation time,intraoperation bleeding volume,postoperative time to first flatus,total hospitalization stay and total hospitalization costs and incidences of postoperative complication were compared.Results The total operation time was (95.0 ±7.0) minutes and (125.0 ± 18.0) minutes,respectively,(P＜0.05).The total costs in the two group were (39515.0 ±4 135.0) yuan and (28287.0 ± 2 254.0) yuan (P ＜ 0.05),respectively.Postoperative complications were observed in 5 (6.1％) and 10 (13.2％) patients (P ＜ 0.05),respectively.The preoperative serum amylase levels were (97.6 ± 48.5) IU/L and (131.4 ± 68.7) IU/L,respectively.The blood loss was (35.7 ± 8.5) ml and (31.8 ± 7.3) ml,respectively.The postoperative time to first flatus was (1.7 ± 0.5) days and (1.9 ± 0.4) days,respectively.The total hospitalization stay was (16.3 ±2.8) days and (15.2 ±3.7) days.There were no significantly differences (P ＞ 0.05).Conclusions LC carried out on day 3 after EST to treat elderly patients with cholecystolithiasis and choledocholithiasis was safe and efficacious and the treatment had the advantages of minimal trauma,short operative time,rapid recovery and low complication rates.This should be recommended in clinical practice.